Smyth M J, Browne K A, Kinnear B F, Trapani J A, Warren H S
Cellular Cytotoxicity Laboratory, Austin Research Institute, Austin Hospital, Heidelberg, Victoria, Australia.
J Leukoc Biol. 1995 Jan;57(1):88-93. doi: 10.1002/jlb.57.1.88.
Cultured natural killer (NK) cells derived from CD3- CD56+ high-density small lymphocytes (HDLs) exhibit similar morphology and high levels of non-major histocompatibility complex-restricted (NK) cytotoxicity equivalent to those of cultured NK cells from CD3- CD56+ low-density large granular lymphocytes (LGLs). To examine the similarities and differences between NK cells from HDLs and NK cells from LGLs, we investigated the expression of three distinct members of the granule serine protease (granzyme) family within cultured CD3- CD56+ LGLs and HDLs. CD3- subpopulations of nonadherent peripheral blood mononuclear cells, LGLs (density < 1.063 g/ml), and HDLs (density > 1.063 g/ml) were stimulated to proliferate in culture. The cultured cells from each population were entirely CD3- CD56+ and were indistinguishable in terms of their increased granularity and size once activated. All cultured CD3- CD56+ LGLs and HDLs displayed cytolytic activity against K562 and immunoglobulin-coated P815. Western analysis detected perforin in both cultured LGL and HDL populations. Cultured HDLs and LGLs both expressed BLT-esterase activity and human granzyme A mRNA. Granzyme B mRNA and protein and Asp-ase activity were detected in unstimulated and cultured LGLs and cultured HDLs. By contrast, unstimulated HDLs did not express significant levels of granzyme B. High levels of Hu-Met-1 granzyme mRNA and Met-ase activity were detected only in cultured LGLs. Thus, despite the development of large granular morphology during proliferation, interleukin-2 cultured CD3- CD56+ HDLs display a different pattern of granzyme expression from CD3- CD56+ LGLs. These data also further suggest an unusually restricted expression of the Hu-Met-1 granzyme in LGLs.
从CD3 - CD56 +高密度小淋巴细胞（HDL）中获得的培养自然杀伤（NK）细胞，其形态与从CD3 - CD56 +低密度大颗粒淋巴细胞（LGL）中获得的培养NK细胞相似，且具有高水平的非主要组织相容性复合体限制（NK）细胞毒性。为了研究HDL来源的NK细胞与LGL来源的NK细胞之间的异同，我们调查了培养的CD3 - CD56 + LGL和HDL中颗粒丝氨酸蛋白酶（颗粒酶）家族三个不同成员的表达情况。对非贴壁外周血单个核细胞的CD3 -亚群、LGL（密度<1.063 g/ml）和HDL（密度>1.063 g/ml）进行刺激使其在培养中增殖。每个群体的培养细胞均完全为CD3 - CD56 +，一旦被激活，其颗粒度和大小增加程度无法区分。所有培养的CD3 - CD56 + LGL和HDL均对K562和免疫球蛋白包被的P815表现出溶细胞活性。蛋白质印迹分析在培养的LGL和HDL群体中均检测到穿孔素。培养的HDL和LGL均表达BLT -酯酶活性和人颗粒酶A mRNA。在未刺激及培养的LGL和培养的HDL中均检测到颗粒酶B mRNA、蛋白质及天冬氨酸酶活性。相比之下，未刺激的HDL未表达显著水平的颗粒酶B。仅在培养的LGL中检测到高水平的Hu - Met - 1颗粒酶mRNA和Met -酶活性。因此，尽管在增殖过程中出现了大颗粒形态，但白细胞介素-2培养的CD3 - CD56 + HDL显示出与CD3 - CD56 + LGL不同的颗粒酶表达模式。这些数据还进一步表明Hu - Met - 1颗粒酶在LGL中表达异常受限。
